人类白细胞抗原基因型的胚胎植入前遗传学诊断  

Research of preimplantation genetic diagnosis of human leukocyte antigen genotype

作　　者：杜生荣[1] 曾国英[1] 孙艳[1] 林运鸿[1] 林典梁[1] 陈清汾 康跃凡[1] Du Shengrong, Zeng Guoying, Sun Yan, Lin Yunhong, Lin Dianliang, Chen Qingfen, Kang Yuefan(Fujian Provincial Maternity and Children＇s Hospital, Affiliated Hospital of Fujian Medical University, Fujian Provincial Reproductive Medicine Center, Fuzhou 350001, Chin)

机构地区：[1]福建省妇幼保健院福建医科大学附属医院福建省生殖医学中心,福州350001

出　　处：《中华生殖与避孕杂志》2018年第8期677-681,共5页Chinese Journal of Reproduction and Contraception

基　　金：福建省计划生育重点专科项目(闽卫科教2012149号);福建省卫生计生青年科研课题(2017-1-17);妇保院研(14-02)~~

摘　　要：目的建立人类白细胞抗原(human leukocyte antigen,HLA)基因配型,为开展HLA配型的胚胎植入前遗传学诊断(preimplantation genetic diagnosis,PGD)联合或不联合单基因病的临床工作提供技术支持。方法采用多重置换扩增(multiple displacement amplification,MDA)的方法扩增3~5个滋养层细胞的全基因组,并用多重聚合酶链式反应(PCR)方法扩增多个短串联重复多态性(polymorphic short tandem repeat,STR)位点,通过毛细管电泳技术验证STR位点的特异性。结果微量细胞全基因组的扩增成功率为93.75%,筛选了HLA基因的5个杂合度高、特异性强的STR位点,等位基因脱扣(allele drop-out,ADO)率为10.34%。结论微量细胞MDA结合多重PCR的方法可以同时检测HLA-A、HLA-B、DRA、DQB位点,扩增成功率高,ADO率低,可以有效地筛选出相匹配的胚胎,为同胞患儿提供造血干细胞来源具有可行性。Objective To provide technical support in preimplantation genetic diagnosis （PGD） of human leukocyte antigen （HLA） matching, associating with or without genetic disease, to establish a multiplex polymerase chain reaction （PCR） for HLA matching in few cells. Methods The whole genome of 3-5 trophoblast cells was firstly amplified with a method of multiple displacement amplification （MDA）. By using the whole genorne as template, the polymorphic short tandem repeat （STR） loci were then amplified with a method of multiplex PCR. The specificity of STR locus was identified with capillary electrophoresis. Results By using this approach, the whole genome was amplified from the few cells, in a rate of 93.75%, and 5 highly heterozygous polymorphic STR loci were screened. The rate of allele drop-out （ADO） was 10.34%. Conclusion Combining the method of MDA in few cells with multiplex PCR, we can simultaneously detect HLA-A, HLA-B, on DRA, DQB loci, and effectively screen the matching embryos to provide child-patients with hematopoietic stem cells in a high amplification success rate and a low ADO rate.

关 键 词：胚胎植入前遗传学诊断(PGD) 人类白细胞抗原(HLA) 多重聚合酶链式反应(PCR) 

分 类 号：R714.8[医药卫生—妇产科学]